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1.
ACS Omega ; 9(14): 16106-16117, 2024 Apr 09.
Article in English | MEDLINE | ID: mdl-38617637

ABSTRACT

The static gravimetric method was used to measure the solubility of 17-α hydroxyprogesterone (OHP) in 13 pure solvents ranging from 278.15 to 323.15 K. The results indicate that the experimental solubility of OHP increases with increasing temperature. The experimental solubility data were correlated by the selected van't Hoff model, λh model, modified Apelblat model, Yaws model, and nonrandom two-liquid (NRTL) model. The fitting results show that the Yaws model can give better correlation results by fitting 13 different pure solvent systems. Based on the NRTL equation, the thermodynamic analysis of solubility data showed that the mixing process was spontaneous. The Hansen solubility parameters (HSPs) and solvent effect were applied to explore these solubility characteristics. Finally, the thermodynamic properties ΔsolH°, ΔsolS°, ΔsolG°, %ξH, and %ξTS were calculated by the van't Hoff model equation. The results showed that ΔsolH°, ΔsolS°, and ΔsolG° are all positive values, indicating that the dissolution of OHP in the selected solvent is an endothermic reaction with increasing entropy.

2.
Zhongguo Dang Dai Er Ke Za Zhi ; 26(1): 48-53, 2024 Jan 15.
Article in Chinese | MEDLINE | ID: mdl-38269459

ABSTRACT

OBJECTIVES: To investigate the clinical characteristics and risk factors of delayed bleeding after intestinal polypectomy in children, and to provide a theoretical basis for clinical surgical intervention of intestinal polyps. METHODS: A retrospective analysis was conducted on the clinical data of 2 456 children with intestinal polyps who underwent endoscopic high-frequency electrocoagulation loop resection in the Endoscopy Center of Children's Hospital Affiliated to Zhengzhou University from January 2014 to December 2021. According to the presence or absence of delayed bleeding after surgery, they were divided into bleeding group with 79 children and non-bleeding group with 2 377 children. A multivariate logistic regression analysis was used to investigate the risk factors for delayed bleeding. The receiver operating characteristic (ROC) curve was used to investigate the value of various indicators in predicting delayed bleeding. RESULTS: Of all 2 456 children, 79 (3.22%) experienced delayed bleeding, among whom 5 children with severe delayed bleeding underwent emergency colonoscopy for hemostasis and 74 received conservative treatment, and successful hemostasis was achieved for all children. There were significant differences between the bleeding and non-bleeding groups in age, body mass index, constipation rate, location of lesion, time of endoscopic procedure, resection method (P<0.05). Children with a diameter of polyps of 6-10 mm and >20 mm were more likely to develop delayed bleeding after resection (P<0.05). The multivariate logistic regression analysis showed that endoscopic operation time, polyp diameter, and resection method were significantly associated with delayed bleeding (P<0.05). The ROC curve analysis showed that the endoscopic operation time, polyp diameter, and resection method had a good value in predicting delayed bleeding after intestinal polypectomy, with an area under the ROC curve of 0.706, 0.688, and 0.627, respectively. CONCLUSIONS: Endoscopic high-frequency electrocoagulation loop resection has a lower incidence of delayed bleeding in children with intestinal polyps, and the endoscopic operation time, polyp diameter, and resection method are closely associated with the occurrence of postoperative delayed bleeding.


Subject(s)
Hemorrhage , Intestines , Child , Humans , Retrospective Studies , Intestinal Polyps/surgery , Risk Factors
3.
ACS Omega ; 8(50): 47463-47471, 2023 Dec 19.
Article in English | MEDLINE | ID: mdl-38144050

ABSTRACT

N-Acetylglucosamine (NAG) is a significant novel functional monosaccharide with a wide range of potential applications, such as in the medical and cosmetic fields. A gravimetric technique was used to assess the solubility of NAG in water, methanol, N,N-dimethylformamide, water + methanol, water + n-propanol, water + N,N-dimethylformamide, and water + acetonitrile at temperatures ranging from 283.15 to 323.15 K. The outcomes of the experiment demonstrated that the temperature and water content were positively correlated with the solubility of NAG in four experimental binary solvents. The order of solubility in the four aqueous solvent mixtures is water + DMF > water + methanol > water + n-propanol > water + acetonitrile. The solubility data was well correlated using the modified Apelblat model, the CNIBS/R-K model, and the Apelblat-Jouyban-Acree model. Experimental data from NAG will help guide the design of cooling and dissolution in crystallization processes.

4.
Cell Commun Signal ; 21(1): 332, 2023 11 20.
Article in English | MEDLINE | ID: mdl-37986081

ABSTRACT

Anemia is the most common manifestation in myelodysplastic syndrome (MDS) patients, but the cause of ineffective hematopoiesis is not fully understood. Enucleation is an important event in the maturation process of erythroblasts. According to a series of morphological phenotypes of the pathological development of MDS erythroblasts, we speculate that there may be enucleation disorders. To verify this hypothesis, we cultured MDS bone marrow CD34+ cells in vitro and induced erythroblast development. The results showed that erythroblast enucleation in MDS was significantly lower than that in the normal group, and the rate of enucleation was positively correlated with hemoglobin concentration. Risk stratification of MDS was performed to further analyze the differences in enucleation among the normal group, low-middle risk group and high-risk group. The results showed that the enucleation rate of the high risk group was higher than that of the low-middle risk group but still lower than that of the normal group. Moreover, the expression of pERK and pAKT in MDS erythroblasts in the high risk group was higher than that in the normal group, while the expression of pERK and pAKT in the low-middle risk group was lower than that in the normal group. Furthermore, the enucleation of MDS was positively correlated with the phosphorylation degree of ERK and AKT. In conclusion, this study reveals that the enucleation of erythroblasts is one of the possible causes of anemia in MDS. Video Abstract.


Subject(s)
Anemia , Myelodysplastic Syndromes , Humans , Erythroblasts/metabolism , Erythroblasts/pathology , Myelodysplastic Syndromes/complications , Myelodysplastic Syndromes/metabolism , Anemia/complications , Anemia/metabolism , Anemia/pathology , Risk Factors , Bone Marrow Cells/pathology
5.
ACS Omega ; 8(42): 39390-39400, 2023 Oct 24.
Article in English | MEDLINE | ID: mdl-37901582

ABSTRACT

The solubility of itraconazole hydroxy isobutyltriazolone (IHI) in four commonly used binary solvent mixtures of N,N-dimethylformamide (DMF) + water, DMF + ethanol, tetrahydrofuran (THF) + water, and THF + ethanol was determined with gravimetric method at temperatures ranging from 283.15 to 323.15 K under atmospheric pressure. The solubility of IHI in all selected solvents increases with the increase of temperature. The maximum solubility of IHI exists in the solvent of DMF + ethanol (0.06523 mol·mol-1, x20 = 0.7, T = 323.15 K), while the minimum solubility exists in DMF + water (0.0003723 mol·mol-1, x20 = 0.3, T = 283.15 K). There is a co-solvency phenomenon in the mixed solvents of DMF+ ethanol, THF + water, and THF + ethanol. Four thermodynamic models, including the modified Apelblat model, the Yaws model, the Sun model, and the modified Jouyban-Acree model, were selected to fit the solubility data of IHI. All the RAD values are less than 0.0484, and RMSD values are not more than 0.001319. The Yaws model and the modified Apelblat model fit the solubility data of IHI better than the other two models. All the selected four models can fit the solubility data of IHI well.

6.
ACS Omega ; 8(28): 25582-25591, 2023 Jul 18.
Article in English | MEDLINE | ID: mdl-37483222

ABSTRACT

The refractive index (RI) is an important physiochemical property of deep eutectic solvents (DESs) for application in the optical identification of specific substances or measuring the concentration of solutes in solutions. However, the available data on the RI of DESs is limited. Here, a systematic investigation on the RI of 48 typical DESs and 30 mixtures with water was conducted under atmospheric pressure. The effect of temperature in the range 293.15-338.15 K, hydrogen-bonding donors (HBDs), and hydrogen-bonding acceptors (HBAs) on RI was investigated. Furthermore, the RI of DESs as a function of mass percentage in the range of 20-80% water was also studied. It was found that the RI of DESs and its aqueous binary mixtures decreases linearly with the increase of temperature. HBDs and HBAs had a significant influence on the RI of DESs. Among them, the RI of choline chloride (ChCl)/phenol and ChCl/o-cresol were obviously higher than those of other DESs. It was also found that the addition of water would decrease the RI of DESs, and the RI of DES content in percentage (wt %) of water binary mixtures increases linearly as a function of mass percentage of DESs for 20 DESs. However, for the other 10 DESs, there is no linear relationship between the RI and the DES content.

7.
Ultrason Sonochem ; 97: 106475, 2023 Jul.
Article in English | MEDLINE | ID: mdl-37321071

ABSTRACT

The objective of this research was to modify the crystal shape and size of poorly water-soluble drug ropivacaine, and to reveal the effects of polymeric additive and ultrasound on crystal nucleation and growth. Ropivacaine often grow as needle-like crystals extended along the a-axis and the shape was hardly controllable by altering solvent types and operating conditions for the crystallization process. We found that ropivacaine crystallized as block-like crystals when polyvinylpyrrolidone (PVP) was used. The control over crystal morphology by the additive was related to crystallization temperature, solute concentration, additive concentration, and molecular weight. SEM and AFM analyses were performed providing insights into crystal growth pattern and cavities on the surface induced by the polymeric additive. In ultrasound-assisted crystallization, the impacts of ultrasonic time, ultrasonic power, and additive concentration were investigated. The particles precipitated at extended ultrasonic time exhibited plate-like crystals with shorter aspect ratio. Combined use of polymeric additive and ultrasound led to rice-shaped crystals, which the average particle size was further decreased. The induction time measurement and single crystal growth experiments were carried out. The results suggested that PVP worked as strong nucleation and growth inhibitor. Molecular dynamics simulation was performed to explore the action mechanism of the polymer. The interaction energies between PVP and crystal faces were calculated, and mobility of the additive with different chain length in crystal-solution system was evaluated by mean square displacement. Based on the study, a possible mechanism for the morphological evolution of ropivacaine crystals assisted by PVP and ultrasound was proposed.

8.
Haematologica ; 108(9): 2487-2502, 2023 09 01.
Article in English | MEDLINE | ID: mdl-37021526

ABSTRACT

Enhancer of zeste homolog 2 (EZH2) is the lysine methyltransferase of polycomb repressive complex 2 (PRC2) that catalyzes H3K27 tri-methylation. Aberrant expression and loss-of-function mutations of EZH2 have been demonstrated to be tightly associated with the pathogenesis of various myeloid malignancies characterized by ineffective erythropoiesis, such as myelodysplastic syndrome (MDS). However, the function and mechanism of EZH2 in human erythropoiesis still remains largely unknown. Here, we demonstrated that EZH2 regulates human erythropoiesis in a stage-specific, dual-function manner by catalyzing histone and non-histone methylation. During the early erythropoiesis, EZH2 deficiency caused cell cycle arrest in the G1 phase, which impaired cell growth and differentiation. Chromatin immunoprecipitation sequencing and RNA sequencing discovered that EZH2 knockdown caused a reduction of H3K27me3 and upregulation of cell cycle proteindependent kinase inhibitors. In contrast, EZH2 deficiency led to the generation of abnormal nuclear cells and impaired enucleation during the terminal erythropoiesis. Interestingly, EZH2 deficiency downregulated the methylation of HSP70 by directly interacting with HSP70. RNA-sequencing analysis revealed that the expression of AURKB was significantly downregulated in response to EZH2 deficiency. Furthermore, treatment with an AURKB inhibitor and small hairpin RNAmediated AURKB knockdown also led to nuclear malformation and decreased enucleation efficiency. These findings strongly suggest that EZH2 regulates terminal erythropoiesis through a HSP70 methylation-AURKB axis. Our findings have implications for improved understanding of ineffective erythropoiesis with EZH2 dysfunction.


Subject(s)
Enhancer of Zeste Homolog 2 Protein , Erythropoiesis , Histones , Humans , Enhancer of Zeste Homolog 2 Protein/genetics , Erythropoiesis/genetics , Histones/metabolism , Methylation , Polycomb Repressive Complex 2/genetics , Polycomb Repressive Complex 2/metabolism
9.
Molecules ; 28(7)2023 Mar 28.
Article in English | MEDLINE | ID: mdl-37049793

ABSTRACT

There are billions of tea drinkers around the world. However, the optimized tea-brewing temperature and time conditions for achieving a higher concentration of antioxidants in tea drinks have not been thoroughly studied. Finding out the optimized brewing conditions can benefit tea drinkers significantly. In this work, we have studied ten antioxidants from seven different popular green, Oolong, black, and scented teas using hot water extraction followed by HPLC analysis. The antioxidant yield was evaluated at 25-100 °C with 5 to 720 min of brewing time. Our results show that the extraction efficiency was enhanced by increasing the water temperature and the highest yield of antioxidants was achieved at 100 °C. The antioxidant yield increased with prolonged brewing time. However, the degradation of antioxidants occurred when tea leaves were extracted for 120 to 720 min. Caffeine was found in all seven tea samples. At 100 °C, the caffein concentration in the tea extract ranged from 7.04 to 20.4 mg/g in Rizhao green tea. Longjing green tea contained the highest concentration of antioxidants (88 mg/g) in the 100 °C extract. Epigallocatechin and caffeine were the most abundant compounds found in all tea samples studied, ranging from 4.77 to 26.88 mg/g. The antioxidant yield was enhanced by increasing the extraction time to up to 60-120 min for all ten compounds studied.


Subject(s)
Antioxidants , Camellia sinensis , Antioxidants/analysis , Caffeine/analysis , Tea , Water , Plant Extracts/analysis
10.
Epigenomics ; 14(20): 1249-1267, 2022 10.
Article in English | MEDLINE | ID: mdl-36420716

ABSTRACT

Aims: To investigate DNA methylation patterns in early and terminal stages of erythropoiesis, and to explore the function of differentially methylated genes in erythropoiesis and erythroid disorders. Materials & methods: Differential analysis of DNA methylation and gene expression during erythropoiesis, as well as weighted gene coexpression network analysis of acute myeloid leukemia was performed. Results: We identified four candidate genes that possessed differential methylation in the promoter regions. DNAJA4 affected proliferation, apoptosis and enucleation during terminal erythropoiesis and was associated with the prognosis of acute myeloid leukemia. DNAJA4 was specifically highly expressed in erythroleukemia and is associated with DNA methylation. Conclusion: DNAJA4 plays a crucial role for erythropoiesis and is regulated via DNA methylation. Dysregulation of DNAJA4 expression is associated with erythroid disorders.


Subject(s)
DNA Methylation , Erythropoiesis , Humans , Erythropoiesis/genetics , Apoptosis , Gene Regulatory Networks , HSP40 Heat-Shock Proteins
11.
ACS Omega ; 7(42): 37663-37673, 2022 Oct 25.
Article in English | MEDLINE | ID: mdl-36312391

ABSTRACT

The solubility data of gliclazide in 10 mono-solvents (1,2-dichloroethane, 1,4-dioxane, 2-methoxyethanol, n-propyl acetate, isopropyl acetate, n-butyl acetate, pentyl acetate, dimethyl sulfoxide (DMSO), N,N-dimethylacetamide (DMA), and 2-butanone) and one kind of binary solvent (DMA + water) were measured between 278.15 and 323.15 K under atmospheric pressure by the gravimetric method. The Hansen solubility parameters and the KAT-LSER equation were used to investigate the solubility order and the influence of solvent effects on solubility. The experimental data were correlated by six thermodynamic models (the λh model, the Yaws model, the Apelblat model, the Jouyban model, the modified Jouyban-Acree model, and the Sun model). The results show that all of these models can correlate the experimental data well. Among them, the Apelblat model is the most suitable for correlating the solubility data of gliclazide in mono-solvents and binary solvents.

12.
Front Immunol ; 13: 972079, 2022.
Article in English | MEDLINE | ID: mdl-36189197

ABSTRACT

Aims: This study aimed to conduct a bibliometric analysis of the relevant literature on the diagnosis of inflammatory bowel disease (IBD), and show its current status, hot spots, and development trends. Methods: The literature on IBD diagnosis was acquired from the Science Citation Index Expanded of the Web of Science Core Collection. Co-occurrence and cooperation relationship analysis of authors, institutions, countries, journals, references, and keywords in the literature were carried out through CiteSpace software and the Online Analysis platform of Literature Metrology. At the same time, the relevant knowledge maps were drawn, and the keywords cluster analysis and emergence analysis were performed. Results: 14,742 related articles were included, showing that the number of articles in this field has increased in recent years. The results showed that PEYRIN-BIROULET L from the University Hospital of Nancy-Brabois was the author with the most cumulative number of articles. The institution with the most articles was Mayo Clin, and the United States was far ahead in the article output and had a dominant role. Keywords analysis showed that there was a total of 818 keywords, which were mainly focused on the research of related diseases caused or coexisted by IBD, such as colorectal cancer and autoimmune diseases, and the diagnosis and treatment methods of IBD. Emerging analysis showed that future research hotspots and trends might be the treatment of IBD and precision medicine. Conclusion: This research was the first bibliometric analysis of publications in the field of IBD diagnosis using visualization software and data information mining, and obtained the current status, hotspots, and development of this field. The future research hotspot might be the precision medicine of IBD, and the mechanism needed to be explored in depth to provide a theoretical basis for its clinical application.


Subject(s)
Bibliometrics , Inflammatory Bowel Diseases , Cluster Analysis , Humans , Inflammatory Bowel Diseases/diagnosis , Machine Learning , Software , United States
13.
Small ; 18(32): e2202663, 2022 08.
Article in English | MEDLINE | ID: mdl-35843879

ABSTRACT

T cell exhaustion caused by mitochondrial dysfunction is the major obstacle of T cells-based cancer immunotherapy. Besides exhausted T cells, the insufficient major histocompatibility complex class I (MHC I) on tumor cells leads to inefficient T cell recognition of tumor cells, compromising therapeutic efficacy. Therapeutic platform to regulate T cell exhaustion and MHC I expression for boosting T cells-based cancer immunotherapy has not been realized up to date. Herein, an injectable hydrogel is designed to simultaneously tune T cell exhaustion and MHC I expression for amplified cancer immunotherapy. The hydrogel is in situ constructed in tumor site by utilizing oxidized sodium alginate-modified tumor cell membrane vesicle (O-TMV) as a gelator, where axitinib is encapsulated in the lipid bilayer of O-TMV while 4-1BB antibody and proprotein convertase subtilisin/kexin type 9 inhibitor PF-06446846 nanoparticles are present in the cavities of hydrogel. After immune response trigged by O-TMV antigen, the 4-1BB antibody-promoted T cell mitochondrial biogenesis and the axitinib-lowered hypoxia synergistically reverse T cell exhaustion while the PF-06446846-amplified MHC I expression facilitates T cell recognition of tumor cells, demonstrating a powerful immunotherapeutic efficacy. This strategy on reprograming T cell exhaustion and improving T cell potency offers new concept for T cells-based cancer immunotherapy.


Subject(s)
Neoplasms , T-Lymphocytes , Antibodies , Axitinib , Histocompatibility Antigens Class I , Humans , Hydrogels , Immunotherapy , Neoplasms/therapy
14.
Acta Biomater ; 147: 366-376, 2022 07 15.
Article in English | MEDLINE | ID: mdl-35588995

ABSTRACT

Immune response is initiated by dendritic cells (DCs), where the cross-presentation of antigens by DCs determines the activating of cytotoxic T cells. However, the efficacy of DCs-initiated immune response is governed by multiple (cascade) steps of immunogenic cell death (ICD), recruitment of DCs, and cross-presentation of DCs. It is urgent but challenging to achieve a platform for simultaneously regulating these multiple steps, amplifying the immune response against tumors. Herein, we reported a photodynamic nanodrug enabling simultaneous regulation of these multiple steps for realizing powerful immune response. The nanodrug was designed by the co-assembling of chlorin e6 (Ce6), celecoxib and 6-thio-2'-deoxyguanosine (6-thio-dG). In our nanodrug, Ce6 enables induction of ICD, while celecoxib down-regulates the prostaglandin E2 (PGE2) for promoting recruitment of DCs enabled by chemokine CCL5 produced from natural killer (NK) cells. Moreover, 6-thio-dG triggers DNA damages in the tumor cells, which in turn activates STING/interferon I pathway for enhancing the cross-presentation ability of DCs. Therefore, an amplified immune therapeutic effect against tumors is achieved, thanks to the simultaneous regulation of these multiple steps. The nanodrug effectively inhibits tumor growth and postoperative recurrence, demonstrating a new approach for boosting immune response initiated by DCs in cancer therapy. STATEMENT OF SIGNIFICANCE: The dendritic cells (DCs)-initiated immune response against tumors is dominated by multiple (cascade) steps including the process of (I) immunogenic cell death (ICD), (II) recruitment of DCs, and (III) cross-presentation of antigens by DCs. Based on this, it is urgent to design a nanoplatform enabling simultaneous regulation of these multiple steps for achieving a potent therapeutic efficacy. A carrier-free photodynamic nanodrug, engineered by a co-assembling approach, was designed to regulate DCs for realizing a powerful DCs-initiated immune response against tumors, thanks to the simultaneous regulation of the above multiple steps. Our nanodrug demonstrated a boosted immune response against tumors, powerfully suppressing primary/abscopal tumor growth and postoperative recurrence, which offers a conceptually innovative strategy for amplifying immunity against tumors.


Subject(s)
Nanoparticles , Neoplasms , Celecoxib , Cell Line, Tumor , Dendritic Cells , Humans , Immunotherapy , Nanoparticles/therapeutic use , Neoplasms/metabolism
15.
ACS Omega ; 7(10): 8591-8600, 2022 Mar 15.
Article in English | MEDLINE | ID: mdl-35309430

ABSTRACT

Metformin hydrochloride (MET·HCl) is one of the most widely used oral hypoglycemic drugs in the world. In addition to hypoglycemic effects, MET·HCl also has anti-inflammatory, anti-tumor, anti-aging, and other effects, showing good efficacy and safety of single and combined treatment. The solubility of MET·HCl in water, water + N,N-dimethylformamide, water + acetonitrile, and water + n-propanol was measured by the gravimetric method under atmospheric pressure at temperatures ranging from 283.15 to 323.15 K. The solubility of MET·HCl has a positive correlation with temperature and water content. The experimental solubility data in binary solvents was correlated by the modified Apelblat model, CNIBS/R-K model, Apelblat-Jouyban-Acree model, and λh model. By comparing the average ARD % values of the four models, it is found that the modified Apelblat model (ARD % = 1.26) provides better correlation. Hansen solubility parameters and apparent thermodynamic parameters were calculated to analyze the solubility behavior, indicating that the dissolution process is endothermic and entropically favorable.

16.
Mikrochim Acta ; 189(4): 145, 2022 03 16.
Article in English | MEDLINE | ID: mdl-35296924

ABSTRACT

A photoelectrochemical (PEC) biosensor based on a multiple signal amplification strategy was established for highly sensitive detection of microRNA (miRNA). TiO2 was prepared on the surface of titanium sheet by laser etching to improve its stability and photoelectrical properties, and CuInS2-sensitized TiO2 was used to form a superior photoelectrical layer, which realized the initial signal amplification. The electron donor dopamine (DA) was modified to H2 as a signal regulator, which effectively increased the photocurrent signal. To further amplify the signal, an enzyme-free hybridization reaction was implemented. When target let-7a and fuel-DNA (F-DNA) were present, the base of H1 specifically recognized let-7a and forced dopamine@AuNPs-H2 away from the electrode surface. Subsequently, the end base of H1 specifically recognized F-DNA, and let-7a was replaced and recycled to participate in the next cycle. Enzyme-free circulation, as a multifunctional amplification method, ensured the recycling of target molecules. This PEC sensor for let-7a detection showed an excellent linear response from 0.5 to 1000 pM with a detection limit of 0.12 pM. The intra-batch RSD was 3.8% and the recovery was 87.74-108.1%. The sensor was further used for clinical biomolecular monitoring of miRNA, showing excellent quantitative detection capability.


Subject(s)
Metal Nanoparticles , MicroRNAs , Nanocomposites , DNA , Dopamine/chemistry , Electrochemical Techniques/methods , Gold/chemistry , Lasers , Limit of Detection , Metal Nanoparticles/chemistry , MicroRNAs/chemistry , Titanium
17.
Blood Adv ; 5(22): 4648-4661, 2021 11 23.
Article in English | MEDLINE | ID: mdl-34551066

ABSTRACT

Enucleation is a key event in mammalian erythropoiesis responsible for the generation of enucleated reticulocytes. Although progress is being made in developing mechanistic understanding of enucleation, our understanding of mechanisms for enucleation is still incomplete. The MAPK pathway plays diverse roles in biological processes, but its role in erythropoiesis has yet to be fully defined. Analysis of RNA-sequencing data revealed that the MAPK pathway is significantly upregulated during human terminal erythroid differentiation. The MAPK pathway consists of 3 major signaling cassettes: MEK/ERK, p38, and JNK. In the present study, we show that among these 3 cassettes, only ERK was significantly upregulated in late-stage human erythroblasts. The increased expression of ERK along with its increased phosphorylation suggests a potential role for ERK activation in enucleation. To explore this hypothesis, we treated sorted populations of human orthochromatic erythroblasts with the MEK/ERK inhibitor U0126 and found that U0126 inhibited enucleation. In contrast, inhibitors of either p38 or JNK had no effect on enucleation. Mechanistically, U0126 selectively inhibited formation/accumulation of cytoplasmic vesicles and endocytosis of the transferrin receptor without affecting chromatin condensation, nuclear polarization, or enucleosome formation. Treatment with vacuolin-1 that induces vacuole formation partially rescued the blockage of enucleation by U0126. Moreover, phosphoproteomic analysis revealed that inactivation of the ERK pathway led to downregulation of the endocytic recycling pathway. Collectively, our findings uncovered a novel role of ERK activation in human erythroblast enucleation by modulating vesicle formation and have implications for understanding anemia associated with defective enucleation.


Subject(s)
Biological Phenomena , MAP Kinase Signaling System , Animals , Cell Differentiation , Endocytosis , Erythroblasts , Humans
18.
Molecules ; 26(13)2021 Jun 30.
Article in English | MEDLINE | ID: mdl-34209151

ABSTRACT

Subcritical water refers to high-temperature and high-pressure water. A unique and useful characteristic of subcritical water is that its polarity can be dramatically decreased with increasing temperature. Therefore, subcritical water can behave similar to methanol or ethanol. This makes subcritical water a green extraction fluid used for a variety of organic species. This review focuses on the subcritical water extraction (SBWE) of natural products. The extracted materials include medicinal and seasoning herbs, vegetables, fruits, food by-products, algae, shrubs, tea leaves, grains, and seeds. A wide range of natural products such as alkaloids, carbohydrates, essential oil, flavonoids, glycosides, lignans, organic acids, polyphenolics, quinones, steroids, and terpenes have been extracted using subcritical water. Various SBWE systems and their advantages and drawbacks have also been discussed in this review. In addition, we have reviewed co-solvents including ethanol, methanol, salts, and ionic liquids used to assist SBWE. Other extraction techniques such as microwave and sonication combined with SBWE are also covered in this review. It is very clear that temperature has the most significant effect on SBWE efficiency, and thus, it can be optimized. The optimal temperature ranges from 130 to 240 °C for extracting the natural products mentioned above. This review can help readers learn more about the SBWE technology, especially for readers with an interest in the field of green extraction of natural products. The major advantage of SBWE of natural products is that water is nontoxic, and therefore, it is more suitable for the extraction of herbs, vegetables, and fruits. Another advantage is that no liquid waste disposal is required after SBWE. Compared with organic solvents, subcritical water not only has advantages in ecology, economy, and safety, but also its density, ion product, and dielectric constant can be adjusted by temperature. These tunable properties allow subcritical water to carry out class selective extractions such as extracting polar compounds at lower temperatures and less polar ingredients at higher temperatures. SBWE can mimic the traditional herbal decoction for preparing herbal medication and with higher extraction efficiency. Since SBWE employs high-temperature and high-pressure, great caution is needed for safe operation. Another challenge for application of SBWE is potential organic degradation under high temperature conditions. We highly recommend conducting analyte stability checks when carrying out SBWE. For analytes with poor SBWE efficiency, a small number of organic modifiers such as ethanol, surfactants, or ionic liquids may be added.


Subject(s)
Biological Products , Hot Temperature , Plant Extracts/chemistry , Sonication , Water/chemistry , Biological Products/chemistry , Biological Products/isolation & purification , Solvents/chemistry
19.
Anal Bioanal Chem ; 413(8): 2207-2215, 2021 Mar.
Article in English | MEDLINE | ID: mdl-33515074

ABSTRACT

We developed a near-infrared (NIR) electrochemiluminescence (ECL) immunosensor for sensitively and selectively determining carbohydrate antigen 125 (CA125) with toxic-element-free and environmental-friendly AgInS2/ZnS nanocrystals (NCs) as tags. The core/shell-structured AgInS2/ZnS NCs not only can be conveniently prepared via an aqueous synthetic procedure, but also has high photoluminescence quantum yield (PLQY) of up to 61.7%, highly monodispersed, water-soluble, and desired biological compatibility. As AgInS2/ZnS NCs can be oxidized via electrochemically injecting holes into their valence band at + 0.84 V, both the monodispersed AgInS2/ZnS NCs in solution and the surface-confined AgInS2/ZnS NCs immobilized in sandwich-typed immuno-complexes with CA125 as analyte can exhibit efficient oxidative-reduction ECL around 695 nm under physiological conditions with the presence of tri-n-propylamine (TPrA). The ECL intensity from the AgInS2/ZnS NCs immobilized in sandwich-typed immuno-complexes increases linearly and selectively with an increased concentration of CA125 from 5 × 10-6 to 5 × 10-3 U/mL, and limit of detection (LOD) was 1 × 10-6 U/mL (S/N = 3). This reliable platform can provide an effective detection method in the early diagnosis and treatment of ovarian cancer.


Subject(s)
CA-125 Antigen/blood , Membrane Proteins/blood , Nanoparticles/chemistry , Silver Compounds/chemistry , Sulfides/chemistry , Zinc Compounds/chemistry , Antibodies, Immobilized/chemistry , Biosensing Techniques/methods , Electrochemical Techniques , Humans , Immunoassay/methods , Limit of Detection , Luminescent Measurements/methods
20.
ACS Omega ; 5(36): 23140-23147, 2020 Sep 15.
Article in English | MEDLINE | ID: mdl-32954164

ABSTRACT

Deep eutectic solvents (DESs) play important roles in the extraction of active constituents in traditional Chinese medicine. Ultrasound-assisted DES has been used to extract flavonoids from Scutellaria baicalensis. Using the contents of scutellarin, baicalin, baicalein, wogonoside, wogonin, and oroxylin A as quantitative indices, different kinds of DESs have been optimized for extraction and betaine/acetic acid has shown the highest yield. The Box-Behnken response surface method (RSM) was utilized to select the extraction conditions with the highest yields. The optimal extraction conditions were as follows: the molar ratio of betaine/acetic acid was 1:4, the water content was 40%, the solid/liquid ratio was 1:100 g/mL, the extraction temperature was 52 °C, and the extraction time was 23 min. Compared with traditional reflux extraction using 70% ethanol as the solvent, ultrasound-assisted DES has a shorter extraction time and higher yields. Furthermore, anti-inflammatory activities of the two extracts by ultrasound-assisted DES and reflux were compared using RAW264.7 cells and the methyl thiazolyl tetrazolium (MTT) method, and they showed equal anti-inflammatory activities. The results demonstrated that the ultrasound-assisted DES method for extraction of flavonoids from scutellariae radix is simple, green, efficient, and reproducible. This research provides good method guides for the rapid and efficient extraction of flavonoids from natural sources.

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